Objective To clone and construct eukaryotic expressing vector of bcr-abl fusion gene and to express the gene in the mammal COS-7 cell lines. Methods bcr-abl fusion gene was amplified from human chronic myeloid leukemia (CML) K562 cell lines by RT-PCR and the fragment of cDNA was retrieved,purified and cloned into the pEGFP-N3 eukaryotic expressing vector. After the selection of the positive clone and by restriction enzyme analysis and DNA sequencing, the correct plasmid was transfected into COS-7 cell lines and observed the transient expression. Results A 874 bp DNA fragment was amplified by RT-PCR. The sequence analysis showed it was consistent with bcr-abl gene of GeneBank. RT-PCR, Western blotting analysis provided strong evidences that bcr-abl gene was expressed successfully in transfected COS-7 cells.Conclusion The eukaryotic expressing vector of bcr-abl fusion gene was constructed, it will lay the foundation for further study of bcr-abl gene in the diagnosis and treatment of CML.

Objective To establish a rapid, specific and sensitive diagnostic technique for the human T. gondii infection with hematopoietic stem cell transplantation and discuss its clinical significance. Me- thods Fifty-six patients subject to hematopoietic stem cell transplantation were detected with ELISA and PCR. Results Among 56 recipients of hematopoietic stem cell transplantation, 7 were positive for T. gondii antigen and 10 were positive for SAG3 gene fragment respectively with the positive rate being 14.3 % and 17.8 % in the ELISA and PCR screening respectively. Twenty healthy people were negative for anti-Toxo antibody.Conclusion PCR is an accurate, relatively rapid, sensitive and specific method for detecting SAG3 gene of T. gondii, and can be considered a valuable additional tool for identification of T. gondii infections.

Objective To investigate IL-18 expression of monocyte cells and the inhibition effect of fluvastation on its expression in patients with acute coronary syndrome(ACS). Methods Isolated monocyte cells from peripheral blood of patients with ACS were incubated with different concentrations of fluvastatin, and IL-18 mRNA level of the monocyte cells was detected by reverse transcription polymerase chain reaction(RT-PCR). Results The monocyte cells of peripheral blood in patients with ACS expressed IL-18 mRNA. IL-18 mRNA decreased from 54 1?9 9 to 50 1?12 6(P

BACKGROUND:Breast cancer stem cel s have a greater impact on the occurrence and metastasis of breast cancer. Under simulated tumor microenvironment, we can better analyze the proliferation and differentiation of breast cancer stem cel s. OBJECTIVE:To explore the tumor microenvironment effect on the differentiation of breast cancer stem cel s. METHODS:Breast cancer cel s and MCF-7 cel s were primarily cultured in fibroblast supernatant and serum-free PCM-2 medium, and formation of breast cancer cel s microspheres was observed. Proliferative ability of breast cancer cel s was detected using MTT colorimetry, and the surface markers of breast cancer stem cel s and epithelial-mesenchymal transition markers were measured using immunocytochemistry and RT-PCR methods. RESULTS AND CONCLUSION:The diameter of primary cel microspheres was larger in the serum-free PCM-2 medium than in the fibroblast supernatant, but the culture speed was faster in the fibroblast supernatant than the serum-free PCM-2 medium. At 3 days of primary culture, the expression of ALDH1 in primary cel s was greatly higher in the serum-free PCM-2 medium than in the fibroblast supernatant. However, the expressions of E-cadherin and vimentin were up-regulated in the fibroblast supernatant than in the serum-free PCM-2 medium. In addition, the expressions of E-cadherin and vimentin in MCF-7 cel s cultured in the fibroblast supernatant were up-regulated, while the expressions of ALDH1 and Oct-4 were downregulated. These findings indicate that the tumor environment has some certain effects on the growth and differentiation of breast cancer stem cel s, and some cytokines secreted from fibroblast supernatant can promote the proliferation and differentiation of breast cancer stem cel microspheres to some extent.

Objective To explore the mechanisms of apoptosis induced by arsenic trioxide and amifostine in human acute promyelocytic leukemia cell lines HL-60 in vitro.Methods HL-60 cells were treated with different concentrations of arsenic trioxide alone and combined with amifostine.The inhibitory ratio of the ceils were measured by MTT assay.and the expression of Survivin Was detected by semiquantitate RT-PCR.Results Proliferation of HL-60 cells exposed to arsenic trioxide dwpped down with increasing dose of the dmg and this effect Was significantly hisher when arsenic trioxide Was used in combination with amifostine.Furthermore.there was a more significant decrease in Survivin expression in HL-60 cells treated with arsenic trioxide in combination with amifostine as compared to the cells treated only with arsenic trioxide.Conclusion Arsenic trioxide induced HL-60 cells to undergo apoptosis by downregulating the expression of Survivin. Amifostine enhanced the sensitivity of HL-60 cells to arsenic trioxide by downregulating the expression of Survivin,thus promoting apoptosis effect.

Objective To investigate the value of interphase fluorescence in situ hybridization(FISH)technique and the detection of fusion gene in the diagnosis of acute myeloid leukemia(AML)M2 and M3 Methods FISH was used to detect the AML1/ETO fusion gene and/or PML/RARα fusion gene in incipient cases including 9 AML-M2, 12 AML-M3 and 10 AML undetermined as AML-M2 or AML-M3 primarily diagnosed by routine morphology though bone marrow,cytochemical staining and immunophenotyping,which can help diagnose and guide clinical therapy.Results 4 of 9 AML-M2 cases were AML1/ETO positive.Among 12 AML-M3 cases,10 were PML/RARα positive.1 case was detected AML1/ETO fusion gene.In 10 untonfirmed M3 or M2,3 case8 showed AML1/ETO,5 showed PMIJRARot fusion gene and the rest showed neither of the genes.Conclusion As a new technique of the molecular genetics,FISH is accurate, rapid and efficient.It would be of significance not only at diagnosis of AML,but also for subsequent clinical decision-making.

AIM: To investigate the protective effects and the mechanisms of prostaglandin E1 (PGE1 ) on ischemia/reperfusion (I/R)-induced lung injury after lung transplantation in rats. METHODS: 36 SD rats were randomly divided into 3 groups ( n = 12 in each) : sham operation group ( control group) , lung transplantation (LT)group and PGE1 treatment group. PGE1 was administered to the rats through intra-venous way from 10 min before the operation to the end of the reperfusion. The wet/dry ratio of lung, lung permeability index and neutro-phil percentage were detected in bronchoalveolar lavage fluid (BALE). Superoxide dismutase (SOD) and malond-ialdehyde (MDA) of lung tissue were measured by color-imetry. Serum level of tumor necrosis factor ?(TNF?)was detected by enzyme-linked immunosorbent assay (ELISA). RESULTS: The wet/dry ratio of lung, lungpermeability index and neutrophils percentage in BALF and MDA content of lung tissue in LT group were higher than those in control group( P

OBJECTIVE: To investigate the effects of arc-track private lock pedicle orthopedic fixation system Ⅱ (ALPF Ⅱ) on treatment of spinal diseases. METHODS: A total of 86 patients with spinal diseases were treated using self-made ALPF in the Wendeng Orthopaedic Hospital and were included in this study. Of these patients, 14 suffered from cervical spinal stenosis complicated by cervical vertebral destabilization, 29 from thoracolumbar fractures and dislocations, 15 from lumbar spinal stenosis complicated by lumbar vertebral destabilization, 2 from lumbar spondylolisthesis, 8 from spinal tuberculosis, 6 from ankylosing spondylitis, 9 from idiopathic scoliosis, and 3 from congenital scoliosis. According to conditions, different therapeutic regimens were selected. Postoperatively, regular follow-up was performed to observe vertebral healing, intervertebral height, spinal column sequence, and neurological function recovery. RESULTS: All patients were followed up for 9-30 months (average 12 months). The improvement rate of neurological function, spinal mobility, back pain, and melosalgia was 94.1%, 65.9%, 92.1%, and 87.4%, respectively. The postoperative anterior and posterior vertebral heights were apparently recovered, and kyphotic angle was well corrected. No screw, rod loosening or breakage was found. CONCLUSION: Self-made ALPF Ⅱ is an internal fixation method for treatment of spinal diseases. It provides good reduction, reliable curative effects, less complications, and no biocompatibilities.

Objective To analyse the fusion genes derived from chromosome structural aberrations in acute myeloid leukemia(AML) and the relationship between fusion genes and the MICM classification, clinical diagnosis, chemotherapy and prognosis. Methods The expression of fusion gene in bone marrow samples was detected with multiplex RT-PCR technique and chromosome karyotypes, immunological phenotypes and clinical data were analyzed in 60 acute myeloid leukemia newly diagnosed. Results 37 cases(61.67 %) of 60 patients carried 5 kinds of fusion genes consisting of MLL-AF9, TLS-ERG, CBFβ-MYH1, AML1-ETO and PML-RARα. The activation of oncogene HOX11 was detected in 13 AML cases, three of them with other chromosome aberration simultaneously.23 cases of 31 patients carrying AML1-ETO or PML-RARα, reached complete remission(CR) after chemotherapy and without relapse. Conclusion Gene typing is the most precise classification method that can direct clinical treatment and evaluate prognosis. Multiplex RT-PCR technique, which can quickly screen 29 kinds of fusion gene derived from chromosome structural aberrations at one time, maybe helpful to improve M1CM classification and guide the choice of treatment.

Objective To explore the deletion of chromosome 13 in multiple myeloma (MM), clinical significance of FISH-defined partial deletion chromosome 13 in MM patients were investigate. Methods Fluorescence in situ hybridization (FISH ) was performed on bone marrow from 38 patients with MM to study the deletion of Rb-1 gene and locus 13q14 on chromosome 13. Fisher exact propability was used to study the relations between partial deletion of chromosome 13 and clinical features. Results 20 out of the 38 cases were found with deletion of chromosome 13; deletion of Rb-1 gene in 4 cases; deletion of locus 13q14 in 2 out of 38 cases; and 14 cases with both of deletions. Fisher exact propability showed that deletion of chromosome 13 was associated with hypso-serum lactic dehydrogenase, stage of ISS. Conclusion Deletion of Rb-1 gene and locus 13q14 were both common cytogenetic changes in MM patients with effect on the biological behavior of the disease, but the value of del (13q14) in MM needs further investigation. FISH was a rapid, accurate and sensitive technique in the analysis of del (13q14) in MM.